LOCUS       sequence-443808-        7595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 gsdA tet-on overexpression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7595)
  AUTHORS   Matthias Steiger
  TITLE     Regulation of the Glucose-6-phosphate dehydrogenase encoding gene 
            gsdA and its impact on growth and citric acid production in 
            Aspergillus niger
  JOURNAL   PLOS One
REFERENCE   2  (bases 1 to 7595)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLOS One"
COMMENT     Sequence Label: pSF8
FEATURES             Location/Qualifiers
     source          1..7595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..73
                     /label=L4440
                     /note="L4440 vector, forward primer"
     intron          812..927
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             join(945..1562,1563..1568,1569..1604,1605..1607,1608..1643,
                     1644..1646,1647..1682,1683..1691)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     protein_bind    join(2039..2057,2058..2080,2081..2099,2100..2122,2123..2141,
                     2142..2164,2165..2183,2184..2206,2207..2225,2226..2248,
                     2249..2267,2268..2290,2291..2309)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     intron          2379..2494
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     terminator      4882..5655
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     promoter        5718..5822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5823..5891,5892..6683)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6041..6060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6909..7497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7398..7417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
       61 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      121 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      181 caacactagt cgcaaaaaac cccgcccctg acagggcggg gttttttcgc ggtctccgat
      241 cggagtcgat aagcttcgga gaatatggag cttcatcgaa tcaccggcag taagcgaagg
      301 agaatgtgaa gccaggggtg tatagccgtc ggcgaaatag catgccatta acctaggtac
      361 agaagtccaa ttgcttccga tctggtaaaa gattcacgag atagtacctt ctccgaagta
      421 ggtagagcga gtacccggcg cgtaagctcc ctaattggcc catccggcat ctgtagggcg
      481 tccaaatatc gtgcctctcc tgctttgccc ggtgtatgaa accggaaagg ccgctcagga
      541 gctggccagc ggcgcagacc gggaacacaa gctggcagtc gacccatccg gtgctctgca
      601 ctcgacctgc tgaggtccct cagtccctgg taggcagctt tgccccgtct gtccgcccgg
      661 tgtgtcggcg gggttgacaa ggtcgttgcg tcagtccaac atttgttgcc atattttcct
      721 gctctcccca ccagctgctc ttttcttttc tctttctttt cccatcttca gtatattcat
      781 cttcccatcc aagaaccttt atttccccta agtaagtact ttgctacatc catactccat
      841 ccttcccatc ccttattcct ttgaaccttt cagttcgagc tttcccactt catcgcagct
      901 tgactaacag ctaccccgct tgagcagaca tcaccgaatt caccatgtct agactggaca
      961 agagcaaagt cataaacggc gctctggaat tactcaatgg agtcggtatc gaaggcctga
     1021 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
     1081 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
     1141 acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
     1201 tccgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
     1261 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
     1321 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
     1381 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
     1441 tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
     1501 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
     1561 gcgggccggc cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     1621 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     1681 tccccgggta actaagtaag gatccactag tacagcagaa gaatctctct ccgctgttgc
     1741 ttcagtgtct gccatgcatt aacttcatcc tactgtccta cccgcagtac ccattcacat
     1801 ttgcggcaga taccaggttg ttttctattc cctcggttct tcagttcttc agatattaca
     1861 ttactcgaga aattggcggc gatgcaggag tttggttggt tttaattgtt tcagtctcct
     1921 tgacctgatt tgtatgacat gcaatgtttc gacggatgac tatctcggtg aatatacgtc
     1981 atgaatcatg cggccgcgcg tatcacgagg ccctttcgac ttcactcgag tttaccactc
     2041 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     2101 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     2161 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
     2221 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
     2281 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcccc atcttcagta
     2341 tattcatctt cccatccaag aacctttatt tcccctaagt aagtactttg ctacatccat
     2401 actccatcct tcccatccct tattcctttg aacctttcag ttcgagcttt cccacttcat
     2461 cgcagcttga ctaacagcta ccccgcttga gcagacatca ccgtttaaac accatggcca
     2521 gcacaatagc acgcactgag gaacgccaga atgctgggtg agttttgcgg tctgcctctc
     2581 tgacatcaca cccctcctcc cactcccgtc cctcctgccc gccgcccaga cgtgaggatt
     2641 caccacccac gactctccat aacaagcccc gtcgcccaaa cattcactgg caggcttccc
     2701 gctttccatt attcttcaat tcgtcaccag gattactcct tcgggcttaa cgaaaggact
     2761 atccttctga ctcaccacca accctcactg cccctcctgc atgctgtagc ggactgcggg
     2821 caccgacctg catcatcgat ctacacccga tccctgtgac attatattcg tcaagctata
     2881 gcctagctaa catggatgtt ttacgtagca ccatggagct caaagatgac actgtcatca
     2941 tagtactggg tgcctccgga gatcttgcaa agaagaaaac cgtcagtgac gaccccctga
     3001 ttcatgttga cctgacagaa agctaacctt ttacagttcc cggccctttt cggccttgta
     3061 tgtcctctcc cagatccaat tgcagtttga ctcaccagta tggttgctga tttgcgcttc
     3121 cagtatcgca acaagttcct ccccaaggga atcaagatcg tcggatatgc ccggacaaac
     3181 atggaccatg aggagtacct gaggcgtgtg cgctcataca tcaagacccc taccaaggaa
     3241 atcgaagagc agctggacag cttctgccag ttctgcacct acatttccgg tcaatatgac
     3301 aaggatgact cgttcatcaa cctcaacaag cacctcgagg agattgagaa gggccagaag
     3361 gagcagaaca gaatctacta catggccctc cctcccagcg ttttcaccac cgtttccgac
     3421 caacttaagc gcaactgcta ccccaagaac ggcgttgccc gtatcatcgt gagtcaatcc
     3481 tgggctggta tcaccctgcc attggtcatt attcttactc gcttgttttc ctatttcaca
     3541 ggtagagaag cctttcggca aggaccttca gagctcgcgc gatctccaaa aagccctgga
     3601 gcctaactgg aaggaagagg agatcttccg tatcgaccac tacctgggta aggagatggt
     3661 caagaacatc cttatcatgc gcttcggaaa cgaattcttc aacgccacct ggaaccgtca
     3721 ccacatcgat aacgttcagg tacgaccttg cgctatccaa ttggcctatt gatttacttg
     3781 ctaaattgtc gcttctatca ttagatcaca ttcaaggagc ccttcggcac tgagggacgt
     3841 ggtggttact tcgatgaatt cggcatcatc cgtgatgtca tgcagaaccg tacgttcaaa
     3901 gtcacgctcg acatctccga catgatgctg ataaaaatct ctcctagacc ttctccaggt
     3961 gttgacgctg ctcgctatgg agcgccccat ttccttctcc gccgaggaca tccgtgacga
     4021 gaaggtacag tgtgcgcttg actattggtt gtgctgggtt actgacactt aaccaggttc
     4081 gtgtcctccg tgcgatggac gccattgagc ccaagaacgt cattattggc cagtacggaa
     4141 agtctctgga tggcagcaag cccgcctaca aggaggacga aaccgttccc caggattccc
     4201 gctgccccac cttctgcgct atggtcgcct acatcaagaa cgagaggtgg gacggtgttc
     4261 ctttcatcat gaaggctggc aagggtatgt acctctttcc aagcgatcat agcaccgatt
     4321 ggtatactaa taattcgcag ccttgaacga gcagaaaacc gagatccgta tccagttccg
     4381 tgacgttacc tccggaattt tcaaggacat ccctcgcaac gagctcgtta tccgcgtcca
     4441 gcccaacgag tccgtgtaca tcaagatgaa ctccaagctg cctggcctgt ccatgcagac
     4501 ggttgtgact gagctcgacc tcacctaccg ccgccgcttc tccgacctca agatccccga
     4561 agcctacgag tctctgatcc tggatgctct gaagggcgac cactccaact tcgtccgtga
     4621 cgatgagctg gatgccagct ggaggatctt cacccctctc ctgcactacc tggatgacaa
     4681 caaggagatc atccccatgg aataccccta cggtacgtgc acttcttgca atttgtctaa
     4741 atcgcttaca tactgaccaa cgcgcaggct cccgcggacc cgccgtcctt gatgacttca
     4801 ccgcgtcctt cggctacaag ttcagcgatg ctgctggcta ccagtggccc ttgacttcca
     4861 cccccaaccg tctgtaagct tgatccactt aacgttactg aaatcatcaa acagcttgac
     4921 gaatctggat ataagatcgt tggtgtcgat gtcagctccg gagttgagac aaatggtgtt
     4981 caggatctcg ataagatacg ttcatttgtc caagcagcaa agagtgcctt ctagtgattt
     5041 aatagctcca tgtcaacaag aataaaacgc gtttcgggtt tacctcttcc agatacagct
     5101 catctgcaat gcattaatgc attggacctc gcaaccctag tacgcccttc aggctccggc
     5161 gaagcagaag aatagcttag cagagtctat tttcattttc gggagacgag atcaagcaga
     5221 tcaacggtcg tcaagagtcc tacgagactg aggaatccgc tcttggctcc acgcgactat
     5281 atatttgtct ctaattgtac tttgacatgc tcctcttctt tactctgata gcttgactat
     5341 gaaaattccg tcaccagccc ctgggttcgc aaagataatt gcactgtttc ttccttgaac
     5401 tctcaagcct acaggacaca cattcatcgt aggtataaac ctcgaaaatc attcctacta
     5461 agatgggtat acaatagtaa ccatgcatgg ttgcctagtg aatgctccgt aacacccaat
     5521 acgccggccg aaactttttt acaactctcc tatgagtcgt ttacccagaa tgcacaggta
     5581 cacttgttta gaggtaatcc ttctttctag aagtcctcgt gtactgtgta agcgcccact
     5641 ccacatctcc actcgctccg gggagaccac cggtgccgct acagggcgcg tcaggtggca
     5701 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5761 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5821 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5881 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5941 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6001 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6061 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6121 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6181 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6241 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6301 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6361 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6421 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6481 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggct
     6541 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6601 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6661 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6721 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6781 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgtaga tctcatgacc
     6841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7561 tttcctgcgt tatcccctga ttctgtggat aaccg
//
